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Methylprednisolone and tacrolimus prevent hypothermia-induced endothelial dysfunction
Antje Diestel1, Nils Billecke, Joerg Roessler
1Department of Pediatric Cardiology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
Insights
Methylprednisolone and tacrolimus protect endothelial cells from hypothermia damage by inhibiting gap formation. This combined drug therapy may prevent endothelial dysfunction after deep hypothermia and rewarming in pediatric cardiac surgery.
Area of Science:
- Cardiovascular Surgery
- Organ Transplantation
- Cellular Biology
Background:
- Hypothermia is a critical method for organ preservation and protecting organs during pediatric cardiac surgery.
- Endothelial cell dysfunction, characterized by tissue edema and function loss, is a common complication in pediatric patients undergoing cardiac surgery and transplantation.
- Investigating protective strategies against hypothermia-induced endothelial damage is crucial for improving patient outcomes.
Purpose of the Study:
- To evaluate the protective effects of methylprednisolone and tacrolimus on endothelial cells subjected to deep hypothermia and rewarming.
- To elucidate the mechanisms by which these drugs modulate endothelial cell function and morphology under hypothermic conditions.
Main Methods:
- Human umbilical vein endothelial cells were pre-treated with methylprednisolone, tacrolimus, or both.
- Cells were exposed to a dynamic cooling and rewarming protocol in a specialized bioreactor or as monolayers.
- Assays included immunocytochemistry, time-lapse microscopy, permeability and adherence tests, and Western blot analysis.
Main Results:
- Hypothermia induced endothelial cell elongation, intercellular gap formation, increased permeability, and reduced adherence.
- Endothelial cell integrity was restored upon rewarming.
- The combined treatment with methylprednisolone and tacrolimus effectively inhibited hypothermia-induced changes, linked to extracellular signal-regulated kinase 1 and 2 (ERK 1/2) activation and connexin 43 expression.
Conclusions:
- Methylprednisolone and tacrolimus prevent hypothermia-induced endothelial gap formation by inhibiting phosphorylated ERK 1/2 and stabilizing connexin 43.
- Combined drug therapy targeting multiple pathways offers a potential strategy to mitigate endothelial dysfunction post-hypothermia.
- This approach may enhance organ preservation and patient recovery in cardiac surgery and transplantation.
Background:
Hypothermia is used to preserve organs for transplantation and is the oldest method to protect organs during complex pediatric cardiac surgery. Loss of tissue function and tissue edema are common complications in children undergoing corrective cardiac surgery and heart transplantation. The present study was designed to examine the effects of methylprednisolone and tacrolimus on endothelial cell function and morphology after deep hypothermia and rewarming.
Methods:
Human umbilical vein endothelial cells were pre-treated with methylprednisolone or tacrolimus, or both, incubated within a specially designed bioreactor or in monolayers, and then exposed to a dynamic cooling and rewarming protocol. Immunocytochemistry, time-lapse video microscopy, cell permeability and adherence assays, and Western blot analysis were performed.
Results:
Confluent endothelial cells exposed to hypothermia displayed elongated cell shapes with intercellular gap formation, increased endothelial cell-layer permeability, and loss in adherence. Upon rewarming, however, endothelial cell integrity was restored. Opening and closing of intercellular gaps was dependent on extracellular signal-regulated kinase 1 and 2 (ERK 1/2) activation and connexin 43 expression. The combined treatment with methylprednisolone and tacrolimus inhibited these hypothermia-induced changes.
Conclusions:
These results suggest that methylprednisolone and tacrolimus inhibit hypothermia-induced endothelial gap formation by phosphorylated ERK 1/2 inhibition and connexin 43 stabilization. Application of combined drugs that affect multiple targets may therefore be considered as a possible new therapeutic strategy to prevent endothelial dysfunction after hypothermia and rewarming.
